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Promoter miR172c and application thereof

A fragment and sequence technology, which is applied to the promoter miR172c and its application fields, can solve the problems of gene expression accumulation in the outer edge, affecting the growth and development of plants, etc., and achieve the effect of strong specific expression activity.

Inactive Publication Date: 2010-09-22
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because its expression does not distinguish between time and space, it will lead to excessive accumulation of outer edge gene expression, which will affect the normal growth and development of plants.

Method used

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  • Promoter miR172c and application thereof
  • Promoter miR172c and application thereof
  • Promoter miR172c and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Preparation of promoter and verification of specific expression

[0025] 1. Preparation and confirmation of the promoter

[0026] 1. Extraction of rice genomic DNA

[0027] (1) Take the leaves of Nipponbare rice, add liquid nitrogen, after fully grinding, pour the powder into a 1.5ml centrifuge tube;

[0028] (2) After the liquid nitrogen in the centrifuge tube is completely volatilized, immediately add 500ul DNA extraction buffer (0.1mol / LTris HCl (pH 8.0), 0.5mol / L NaCl, 0.05mol / L EDTA, 0.5% SDS), fully After mixing, incubate at 65°C for 30 minutes, during which the centrifuge tube is gently inverted up and down to fully mix the sample with the buffer;

[0029] (3) Centrifuge at room temperature for 15 minutes at 12,000 rpm;

[0030] (4) Transfer the supernatant to another centrifuge tube;

[0031] (5) Add an equal volume of tris-phenol and extract once;

[0032] (6) Extract again with equal volume of chloroform;

[0033] (7) Take the supernatant, add...

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Abstract

The invention discloses a promoter miR172c and application thereof. The promoter is a DNA fragment as shown in 1), 2), 3) or 4), i.e. 1), a DNA fragment as shown in sequence 1 in a sequence table; 2), a DNA fragment hybridized with the DNA sequence limited by 1) under strict conditions and haviing the same function with the DNA fragment shown in 1); and 3), a DNA fragment having more than 90% homology with the DNA sequence limited by 1) and the same function with the DNA fragment shown in 1). Experiments prove that the miR172c DNA fragment in the invention is a promoter, and the promoter has strong peculiar expression activity, and is peculiarly expressed in the stele of the root of a plant. The promoter provides useful means for researches on functions of genes peculiarly expressed and transgenosis breeding of plants, and has important meanings.

Description

technical field [0001] The present invention relates to a promoter miR172c and its application. Background technique [0002] Promoters can be divided into three categories according to their mode of action and function: constitutive promoters, inducible promoters, and tissue-specific promoters. Constitutive promoters can initiate expression in all tissues, with continuous expression and no time-space specificity, such as the CaMV35S promoter. Such promoters have been extensively studied and applied, and play an important role in genetic engineering. However, because its expression does not distinguish between time and space, it will lead to excessive accumulation of outer edge gene expression, which will affect the normal growth and development of plants. Therefore, searching for inducible or tissue-specific promoters plays an important role in gene function research and transgenic breeding. Contents of the invention [0003] One object of the present invention is to p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00
Inventor 王秀杰王猛储成才曹守云
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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